Non-fouling polymeric surface modification and signal amplification method for biomolecular detection

• US 9,493,823 B2
• Filed: 01/03/2013
• Issued: 11/15/2016
• Est. Priority Date: 09/15/2005
• Status: Active Grant

• Current Assignee: Duke University
• Original Assignee: Duke University
• Inventors: Chilkoti, Ashutosh
• Primary Examiner(s): Lam, Ann
• Application Number:
  US13/733,302
• Publication Number:
  US 20130143771A1
• Time in Patent Office: 1,412 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00432 Photolithographic masks
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00628 Ionic
• CPC Class Codes: B01J 2219/0063 Other, e.g. van der Waals forces, hydrogen bonding
• CPC Class Codes: B01J 2219/00635 by reactive plasma treatment
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00677 Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01J 2219/0074 Biological products
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C12Q 1/682 Signal amplification
• CPC Class Codes: C12Q 1/6837 : using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 2521/131 : Terminal transferase
• CPC Class Codes: C12Q 2525/313 : Branched oligonucleotides
• CPC Class Codes: C40B 50/18 : using a particular method of attachment to the solid support
• CPC Class Codes: G01N 33/5302 : Apparatus specially adapted for immunological test procedures
• CPC Class Codes: G01N 33/54353 : with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: G01N 33/54393 : Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding

Abstract

An article such as a biosensor having a nonfouling surface thereon is described. The article comprises: (a) a substrate having a surface portion; (b) a linking layer on the surface portion; (c) a polymer layer comprising brush molecules formed on the linking layer; and (d) optionally but preferably, a first member of a specific binding pair (e.g., a protein, peptide, antibody, nucleic acid, etc.) coupled to the brush molecules. The polymer layer is preferably formed by the process of surface-initiated polymerization (SIP) of monomeric units thereon. Preferably, each of the monomeric units comprises a monomer (for example, a vinyl monomer) core group having at least one protein-resistant head group coupled thereto, to thereby form the brush molecule on the surface portion. Methods of using the articles are also described.

Claims

1. A biomolecular detector, comprising:

(a) a substrate having a surface portion;

(b) a linking layer on said surface portion; and

(c) a polymer layer formed on said linking layer by a process of surface-initiated polymerization of monomeric units thereon, with each of said monomeric units comprising a monomer core group having at least one protein-resistant head group coupled thereto, to thereby form a brush molecule on said surface portion;

said brush molecule comprising a stem formed from the polymerization of said monomer core groups, and a plurality of branches formed from said head group projecting from said stem; and

(d) a first member of a specific binding pair coupled to said brush molecule.

2. The detector of claim 1, wherein said member of a specific binding pair further comprises an extended nucleic acid conjugated thereto, said extended nucleic acid produced by a process of enzymatic extension with terminal transferase.

3. The detector of claim 1, wherein said first member is a first nucleic acid, said detector optionally further comprising a second nucleic acid as a second member of said binding pair hybridized to said first member.

4. The detector of claim 3, further comprising an extended nucleic acid produced by a process of enzymatic extension with terminal transferase coupled to either said first or second nucleic acid.

5. The detector of claim 1, wherein said member of a specific binding pair is a protein or peptide.

6. The detector of claim 5, further comprising:

a second member of said specific binding pair coupled to said first member of said specific binding pair;

an antibody specifically bound to said second member to form a sandwich, said antibody having a first nucleic acid conjugated thereto; and

an extended nucleotide coupled to said first nucleotide, said extended nucleic acid produced by a process of enzymatic extension with terminal transferase.

7. The detector of claim 1, wherein said first member is an antibody.

8. The detector of claim 1, wherein said surface portion comprises a material selected from the group consisting of metals, metal oxides, semiconductors, polymers, silicon, silicon oxide, and composites thereof.

9. The detector of claim 1, wherein said surface portion comprises gold.

10. The detector of claim 1, wherein said linking layer is continuous.

11. The detector of claim 1, wherein said linking layer is patterned.

12. The detector of claim 1, wherein said linking layer is a self-assembled monolayer.

13. The detector of claim 1, wherein said brush molecule is from 5 to 50 nanometers in length.

14. The detector of claim 1, wherein said brush molecule is formed on said surface portion at a density from 40 to 100 milligrams per meter₂.

15. The detector of claim 1, wherein said brush molecule is produced by surface-initiated atom transfer radical polymerization of oligo(ethylene glycol)methyl methacrylate.

16. The detector of claim 1, wherein said head group comprises a kosmotrope.

17. The detector of claim 1, wherein said first member of said specific binding pair is coupled directly to a hydroxyl group in OEGMA by CDI chemistry.

18. A biomolecular detector, comprising:

(a) a substrate having a surface portion;

(b) a linking layer on said surface portion; and

(c) a polymer layer formed on said linking layer by a process of surface-initiated polymerization of monomeric units thereon, with each of said monomeric units comprising a monomer core group having at least one protein-resistant head group coupled thereto, to thereby form a brush molecule on said surface portion;

said brush molecule comprising a stem formed from the polymerization of said monomer core groups, and a plurality of branches formed from said head group projecting from said stem; and

(d) a first nucleic acid as a first member of a specific binding pair coupled to said brush molecule.

19. The detector of claim 18, wherein said surface portion comprises a material selected from the group consisting of polymers, silicon, silicon oxide, and composites thereof.

20. The detector of claim 18, wherein said linking layer is continuous.